Ensemble machine learning with Bayesian optimization predicts bioactive extraction from fermented watermelon rind...

A new integrated extraction strategy was established by combining natural solvent systems (NADES), microwave-assisted extraction, and machine learning techniques for optimizing the extraction of bioactive compounds from fermented watermelon rind. The results showed that solid-state fermentation significantly improved extraction rates, compared to non-fermented controls. The fermentation pretreatment comparison was conducted under identical extraction conditions, with non-fermented watermelon rind serving as the internal control. Machine learning models optimized using Bayesian optimization were developed to describe and predict three important response variables: TPC, TFC, and antioxidant activity, as functions of four extraction variables (microwave power, temperature, time, and solid-liquid ratio). The ensemble models showed outstanding predictive capabilities with test R² values of 0.9147, 0.9088, and 0.9252 for TFC, TPC, and DPPH activity, respectively, with very low overfitting (ΔR² < 0.06). Importance analysis of the features showed temperature as the most important parameter in the extraction of bioactive compounds (importance: 0.842-0.885), followed by solid-liquid ratio. Simultaneous optimization using the validated models showed the optimal extraction conditions to be 62.5 °C, 27.7 min, 300 W, and 30 mg/mL, predicting values of 1.656 mg CE/g (TFC), 19.80 mg GAE/g (TPC), and 71.27% (DPPH activity). Solid-state fermentation enhanced extraction yields, increasing total phenolics (16.9 to 19.1 mg GAE/g), flavonoids (0.61 to 0.74 mg CE/g), and DPPH activity (66% to 75%). The developed ensemble models showed high accuracy (R² = 0.91-0.93; RMSE = 0.0812 mg/g, 0.51 mg/g, and 0.74%). Experimental validation results have confirmed the high accuracy of the model with relative errors of less than 3% for all responses. The combination of fermentation pretreatment, green extraction, and machine learning indicates promise for sustainable valorization of agricultural waste resources as sources of bioactive compounds for nutraceutical applications.